The strain of bacteria desulfovibrio vulgaris east-1, used for the treatment of wastewater from heavy metal ions

 

(57) Abstract:

The invention relates to industrial Microbiology and concerns of biotechnology for clean industrial wastewater. A new strain of bacteria Desulfovibrio Vulgaris EAST-1 is proposed for the treatment of wastewater from heavy metal ions. The strain is highly resistant in growth in environments contaminated with metal ions, provides a stable biomass growth. The technical result of the invention is an effective treatment of wastewater by binding and removing heavy metal ions. table 1.

The invention relates to industrial Microbiology and relates to a new strain of bacteria - producers of sulfide, which can be used in biotechnology for clean industrial wastewater engineering, instrument engineering, electrical engineering enterprises.

Aware of the use of sulfate reducing bacteria (SRBs) for the treatment of wastewater from heavy metal ions. During biochemical purification of the binding of heavy metal ions biogenic sulfide in insoluble sulfides, as well as the recovery of some metals and precipitation as hydroxides. To improve the efficiency of methods of biochemic the project required concentration of microorganisms in the sewage treatment process from ions of heavy metals using different techniques or add easily digestible organic food for SSC [1] or use specially selected communities of microorganisms [2].

However, the applied microorganisms do not provide special growth activity of sulfate reducing bacteria in industrial wastewater, characterized by different degree of pollution and different composition of contaminants. Adopted as a prototype strain of bacteria Desulfovibrio Vulgaris BKM B-1970 D (UV-3), used for treatment of wastewater from heavy metal ions. This strain is highly active strains of sulfate-reducing bacteria, providing a stable growth of biomass in waste water from heavy metal ions [3].

The strain of sulfate reducing bacteria Desulfovibrio Vulgaris BKM B-1970 D obtained by extraction from wastewater electroplating with a high content of dissolved metals and the subsequent selection of standard methods for the elective environments. The main composition of the medium shown in example.

The resulting strain of bacteria deposited in the all-Union collection of microorganisms (BKM) number BKM B-1970 D.

The definition of culture given by the determinant of Berga ("Bergey''s Manual of Determinative Bacteriology", v. 1, 909, Baltimore William, Wilkins, 1984).

Example. Cultural and morphological characteristics of the strain.

Characterization of vegetative cells, the vibrios, the cell size of 0.4 to 0.7; 1.5 to 4 microns; gram-negative; my uniform division, type jgutikova - motile, polar jgutikova; on no. Type colonies - colonies are spherical and discoidal in the thickness of the agar, growth characteristics on nutrient - restore sulphate formation of sulfides.

Physiological and biochemical characteristics of the strain.

Conditions for the cultivation of anaerobic, the optimum temperature is 30oC (within 10-35oC), the optimum pH value of 7.2 to 7.4 (range 5.5 to 8.5), the change in pH during growth is absent or weak alkalization. The smell is hydrogen sulfide.

Relation to pH, temperature and oxygen specified in the conditions of cultivation, is inhibited by streptomycin, ampicillin and other antibiotics, any abscopal gram-negative microflora. Cells do not contain phages, not lysogeny, maintain the salt content of up to 20-30 g/l, gelatin liquefies, hydrolyzes starch is not.

This strain was used for the purification of waste water of electroplating.

The disadvantage of this strain is not sufficiently high efficiency wastewater treatment electroplating plants from heavy metal ions.

The objective of the invention is obtaining a strain of bacteria Desulfovibrio Vulgaris BKM B-1020 D, use ERI, used for wastewater treatment. The essence of the invention is achieved by applying a strain of bacteria Desulfovibrio Vulgaris BKM B-1020 D for wastewater treatment galvanic production. Bacterial strain isolated from the coolant (coolant) metalloobrabatyvajushhego shop.

The proposed strain deposited at the all-Union collection of microorganisms (BKM) under registration number - BKM B-2010 D.

The proposed strain is characterized by the following morphological-cultural, physiological and biochemical properties.

1. Cultural and morphological features of patentable strain is defined in its cultivation on standard environments, the recipe of which is taken from the catalogue of the all-Russian collection of microorganisms 92 and 109 and following the publication of "all-Russian collection of microorganisms. Directory", Pushchino-Moscow technological Institute, 1992, 362 pages Cultivation was performed under anaerobic conditions at 30oC for 2 days.

Cultural characteristics: the growth of strain liquid medium opalestiruet, at the bottom of the vessel cultivation of accumulated black precipitate of sulphide of iron; the smell of hydrogen sulfide.

Morphological features: microscopic examination of cells presents dinoguy (monotreme); multiplication division with a uniform divergence of cells. By building the wall - gracilicutes, as evidenced by a negative gram stain. Not acid-fast. Spores do not form. Intracellular inclusions under cultivation in these environments are missing.

Growth characteristics: recovery of sulfate to sulfide (hydrogen sulfide) with the above mentioned accumulation in standard black iron sulfide, and the exhaustion of iron) of hydrogen sulfide.

2. Physiological and biochemical characteristics of patentable strain

Strict gone anaerobic. Capable of sulfate respiration: as electron acceptor uses sulfate or sulfur compounds of intermediate oxidation (sulfite, thiosulfate) as donors of hydrogen using: hydrogen, lactate, pyruvate, formate, ethanol. Respectively as sources of carbon are able to use the above named lactate, pyruvate, formate, ethanol. In addition, can be used as a carbon source (growth substrate) industrial coolant. As nitrogen sources can use ammonium and amino acids yeast extract. The gelatin liquefies, hydrolyzes starch is not. In stimulants releasee with the substitution of sulphate sulphide. The temperature range for growth of strain 0-41oC with an optimum at 3oC. Cells contain cytochrome C and desulfoviridin. Growth of the strain is inhibited by streptomycin, ampicillin and other antibiotics, any abscopal Gram-negative flora.

3. Recipe environments from the catalogue-Russian collection of microorganisms.

N 92. Medium with lactate for DESULFOVIBRIO

A solution of 1 g:

K2HPO4- 0.5

NH4Cl - 1.0

CaCl26H2O - 0.1

MgSO47H2O - 2.0

Na2SO4- 1.0

Lactate Na - 5.0

Yeast extract - 1.0

Resazurin - 0.001

Cysteine - 0.5

Distilled water, ml - 950,0

Solution 2.

NaHCO3, g - 4

Distilled water, ml - 40

Solution 3.

Na2S9H2O, g - 0.3

Distilled water is 6.0

Solution 4.

FeSO47H2O, g - 0.4

Distilled water, ml - 10.0

Preparation: solution 1 bring to boil with simultaneous blowing through the solution free from oxygen gas mixture consisting of 97% N2and 3% H2. Sterilize in the atmosphere of these gases. A solution of 3 sterilized in an atmosphere of N2. After sterilization solutions to connect. the pH of the medium after see the BR>
NH4Cl - 1.0

Na2SO4- 1.0

CaCl26H2O - 0.1

MgSO47H2O - 2.0

Lactate Na - 2.0

Yeast extract - 1.0

Resazurin - 1.0

Distilled water, ml - 980.0

Solution 2.

FeSO47H2O, g - 0.5

Distilled water, ml - 10.0

Solution 3.

Thioglycolate Na, g - 0.1

Ascorbic acid, 0.1

Distilled water, ml - 10.0

A solution of 4 (trace element solution SL-10), mg:

HCl(25%, 7.7 M), ml - 10.0

FeCl24H2O, g - 1.5

ZnCl2- 70.0

MnCl24H2O - 100.0

H3BO3- 6.0

CoCl26H2O - 190.0

CuCl22H2O - 2.0

NiCl26H2O - 24.0

Na2MoO42H2O - 36.0

Distilled water, ml - 990.0

Preparation: all the solutions are sterilized separately by autoclaving at 121oC for 15 min in nitrogen atmosphere. Solution 1 immediately before autoclaving boil, blowing with nitrogen. To prepare the environment mix solutions 1, 2 and 3 and solution 4 was added 1 ml, pH of the medium after mixing set 7,2-7,4.

Example 1. Obtaining strain Desulfovibrio Vulgaris BKM B-2010 D

The deposited strain has been obtained by sampling the manufacture of the 30oC on Wednesday, the recipe of which is given in the text, with the modification of environments that as the sole carbon source for growth production use coolant. Then the selection of the strain continue with increasing concentrations of heavy metals in the environment of the cultivation. The final isolation of pure culture of patentable strain produced by the serial dilution method with successive planting in agar option nutrient medium and subculture of single colonies.

An example of using a strain of bacteria Desulfovibrio Vulgaris BKM B-1020 D for the treatment of wastewater from heavy metal ions.

The process is carried out in biotene downloading pumice with upward flow of the liquid. Seeds are grown in liquid medium in flasks strains of microorganisms. Sow biotene microorganisms Desulfovibrio Vulgaris BKM B-1020 D, which are fixed on the pumice. The temperature of cultivation in biotene support 28-30oC. Through biotank pass the waste water of electroplating. As a result of life SSC released hydrogen sulfide, which is connected with heavy metal ions, forming sulfides of metals that precipitate.

The effectiveness of the work of the economic production using the proposed strain is 99.9%.

Thus, the waste water of electroplating plants can be effectively cleaned from heavy metal ions using strain Desulfovibrio Vulgaris BKM B-1020 D (EAST-1).

LITERATURE

1. USSR author's certificate 857013, class C 02 F 3/28, 1979

2. USSR author's certificate 941319, class C 02 F 3/34, 1980

3. RF patent N 2055817, class C 02 F 3/34, 1996

The strain of bacteria Desulfovibrio Vulgaris WCMW-2010 D used for treatment of wastewater from heavy metal ions.

 

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5 cl, 8 dwg, 10 ex

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